neighboring cells . As major trans-membrane proteins, CLDNs play crucial roles in the formation and maintenance of tight junctions . It is generally accepted that the disruption of tight junctions leads to the loss of intercellular cohesion, which contributes to the invasiveness and lack of differentiation of cancer cells and thus promotes metastasis. Earlier studies have suggested that mRNA or membrane protein expression levels of CLDNs were strongly correlated with carcinogenesis in BC and especially CLDN1 [13–19]. However, clinical studies are still relatively limited. Two reports have suggested a correlation between CLDN1 down-regulation and BC recurrence [16,17]. One study reported that down-regulation of CLDN2 was associated with advanced BC . Of the few publications on CLDN7 expression in BC, researchers have reported that positive CLDN7 expression was significantly associated with an increased risk of recurrence and nodal involvement but with lower histological grade in a small sample of invasive ductal carcinoma (IDC) tumors [19,20]. The roles of the four CLDNs listed above are not well understood with respect to the various subtypes of TNBC.
of mammary stem cells and Wnt/beta-catenin signaling pathway. Lin SY and coworkers reported the nuclear staining of beta- catenin was associated with poor outcome of breastcancer patients . However, several other studies failed to find such association [24,25]. One possible explanation is these studies did not categorize the beta-catenin localization by molecular subtypes. Khramtsov AI and colleagues previously reported that Wnt/beta- catenin pathway activation was enriched in basal-like breast cancers . There are significant limitations to this current study, including the small data set, few cases of stage I patients, no validation in an independent series of cases, categorize the accumulation of DKK1 and beta-catenin inbreast cancers by immunohistochemical to positive and negative rather than scoring. Despite these shortcomings, our results do support the idea that Wnt/beta-catenin signaling pathway is activated intriplenegativebreast cancers.
Introduction: Breastcancer is the most common visceral malignancy in women, the leading cause of cancer death among women worldwide. The triplenegative subgroup has poorprognosis and aggressive biological behavior. Objectives: To outline the clinical and histopathological aspects, the treatment proile, and to suggest which factors may predict poorprognosisin patients with triple-negative invasive breastcancerin the Campos Gerais region of Paraná. Methods: A retrospective observational cohort study, longitudinal, comparative, performed in a clinic of anatomic pathology in the Instituto Sul Paranaense de Oncologia, in Ponta Grossa, Paraná. The inclusion criteria were female patients with pathology report of invasive breast carcinoma, whose immunohistochemistry showed negative for hormone receptors and human epidermal growth factor receptor (HER2), diagnosed in the period between January 1, 2002 and December 31, 2012. The patients were divided into two groups, living women and patients who have died. Results: The recurrence rate, chemotherapy type, angiolymphatic invasion, tumor size, lymph node invasion, and type of surgery performed were signiicant variables in the univariate analysis between the groups. After Cox regression for multivariate analysis, only the angiolymphatic invasion (p = 0.012, relative risk [RR] 5.0518, conidence interval [CI] 95% 1.4261-17.8952), and tumor size (p = 0.0385, RR 1.2605, CI 95% 1.0123-1.5695) remained signiicant. Conclusion: The angiolymphatic invasion and tumor size proved to be risk factors for death, from all causes, in patients with triple-negativebreastcancer. Differences between groups can indicate different molecular subtypes within the triple-negativephenotype.
In our previous study  we had used unsupervised methods to identify subgroups of TNBC without considering outcome in the first place. Based on subsequent correlation of the obtained groups with prognosis we then constructed a simple binary classifier from expression of B-cell- and IL-8-metagenes. In contrast, the supervised signature presented here seem to include information from several additional biological characteristics. In fact this supervised signature can outperform the simple combination of the two parameters used in our previous study. However, the interpretation of the biology of such an amalgamated signature could be much more difficult than the interpretation of metagenes. In summary, in this paper we demonstrated that the use of a homogenous TNBC dataset allowed us to identify prognostic gene signatures that are unrelated to previously published general breastcancer prognostic signatures. The composition of the signature suggests that IL-8 mediated inflammation and VEGF related signaling herald very poorprognosisin TNBC and immune infiltration predicts better outcome. These observations could also suggest potential novel therapeutic strategies for these patients as e.g. inhibiting IL-8 signalling [51,52] might be combined with anti-angiogenesis therapies , and immune augmentation .
LSD1, a lysine-specific histone demethylase, is overexpressed in several types of cancers and linked to poor outcomes. Inbreastcancer, the significance of LSD1 overexpression is not clear. We have performed an in silico analysis to assess the relationship of LSD1 expression to clinical outcome. We demonstrate that LSD1 overexpression is a poor prognostic factor inbreastcancer, especially in basal-like breastcancer, a subtype of breastcancer with aggres- sive clinical features. This link is also observed in samples of triplenegativebreastcancer. In- terestingly, we note that overexpression of LSD1 correlates with down-regulation of BRCA1 intriplenegativebreastcancer. This phenomenon is also observed inin vitro models of basal- like breastcancer, and is associated with an increased sensitivity to PARP inhibitors. We pro- pose therefore that high expression levels of the demethylase LSD1 is a potential prognostic factor of poor outcome in basal-like breastcancer, and that PARP inhibition may be a thera- peutic strategy of interest in this poor prognostic subtype with overexpression of LSD1.
phenotype and for the selection of the most efficient therapies (Weigelt and Reis-Filho 2010). Triple-negativebreastcancer (TNBC) is a subtype characterized by the lack of ER, PR, and HER2 expression and is associated with younger age at diagnosis (Dent et al. 2007) and occurs with greater frequency in premenopausal African- American women (Carey et al. 2006). It represents approximately 12–17% of all breast cancers (Foulkes et al. 2010) and encompasses a heterogeneous group of tumors, including, but not limited to, those classified as basal-like. There is an unmet need to better understand the drivers of this breastcancer subtype because the usual antiendocrine and anti-HER2 targeted therapies are ineffective, and traditional cytotoxic chemotherapy seems to be insufficient (Cadoo et al. 2013). The aggressive clinical course, poorprognosis, and lack of specific therapeutic options have intensified the current interest in this subtype of tumor (Cho et al. 2011).
Triple-negativebreastcancer (TNBC) is defined by the lack of the expression of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2). It is characterized by aggressive behavior, poorprognosis and lack of targeted therapies. MicroRNA (miRNA) as a novel modulator of gene expression has played an important regulatory role in the malignancy. Dysregulation and/or mutation of the miRNAs may also contribute to the TNBC susceptibility since it is associated with the expression of ER, PR and HER2. Single nucleotide polymorphisms (SNPs) in miRNAs may be extremely relevant for TNBC. We tried to validate the hypothesis that genetic variations in miRNA are associated with TNBC development, and identify candidate biomarkers for TNBC susceptibility and clinical treatment. We screened the genetic variants in all miRNA genes listed in the public database miRBase and NCBI. A total of 23 common SNPs in 22 miRNAs, which tagged the known common variants in the Chinese Han people with a minor allele frequency greater than 0.05, were genotyped. This case-control study involved 191 patients with TNBC and 192 healthy female controls. Frequencies of SNPs were compared between cases and controls to identify the SNPs associated with TNBC susceptibility. No significant association was found between TNBC risk and the SNPs in the miRNA genes in the Chinese Han people (P.0.05), but this warrants further studies.
Ki-67 is a non-histone nuclear protein that is closely linked to proliferation cells. High Ki-67 expression is associated with a higher histologic grade, larger tumor size, the presence of axillary lymph nodal metastasis, and worse outcome[21–23]. In neoadjuvant chemotherapy (NAC), higher Ki-67 expression is associated with better pathologic complete response (pCR); however, it is a strong prognostic factor for worse survival of patients who fail to achieve pCR after NAC[24–26], suggesting that Ki-67 can be used for classification of patients with different responses and prognosis. Ki-67 is one of the markers for chemosensitivity inbreast carcinomas, but the correlation between Ki-67 expression and chemosensitivity in the triple- negativephenotype is not clear, probably due to heterogeneous characteristics of TNBC. Furthermore, standardized staining methods and image analysis for making a consistent cut-off value for Ki-67 may be required to achieve a comparative measurement[10,27]. Our study indicates that a cut-off level of 20% for Ki-67 index is adequate to demonstrate a correlation with recurrence.
TNBC is generally associated with a poor outcome, which is essentially not predicted by assessment of standard clinico- pathological variables, such as lymph node status or tumour size at initial presentation. The lack of identified molecular targets in the majority of TNBCs implies that chemotherapy remains the treatment of choice for patients with TNBCs. Here we show that, regardless of the reason that led to an absence of adjuvant therapy for patients involved in the GSE31519 study, those with a high macroH2A1.1/macroH2A1 mRNA ratio have a worse prognosis than those with a low one. Even if this observation clearly needs to be confirmed with a larger cohort, it is tempting to propose that assessing macroH2A1.1 expression levels will allow the identifica- tion of TNBC patients who, despite favorable clinic-pathological variables such as lymph node status or tumour size at initial presentation, will have a worse prognosis and may benefit from
Increased levels of Skp2 and reduced levels of p27 occur in various types of cancer, such as gastric carcinoma , prostate cancer , oral squamous cell carcinoma , and diffuse large B- cell lymphoma . Inbreastcancer, Zheng et al. reported that high level of Skp2 expression were more frequently found in ER- negative tumors and tumors with metastatic axillary lymph nodes . Traub et al. found that the combined assessment of Skp2 and p27 expression identifies aggressive breastcancer, and high Skp2 and low p27 expression indicates an unfavorable clinical course . All the above mentioned studies analyzed the relationship of nuclear Skp2 expression with clinicopathological characteristics, concluding that nuclear Skp2 expression predicted a poorprognosis.
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21. Bear HD, Anderson S, Brown A, Smith R, Mamounas EP, Fisher B, Margolese R, Theoret H, Soran A, Wickerham DL, Wolmark N; National Surgical Adjuvant Breast and Bowel Project Protocol B-27. The effect on tumor response of adding sequential preoperative docetaxel to preoperative doxorubicin and cyclophosphamide: preliminary results from National Surgical Adjuvant Breast and Bowel Project Protocol B-27. J Clin Oncol. 2003;21(22):4165-74. 22. von Minckwitz G, Schneeweiss A, Salat C, Rezai M, Zahm DM, Klare
To determine the prognostic value of RANKL/RANK expression, survival analysis was performed. All patients were followed up until July 2014. Among the total 116 patients, 66 patients (57%) died during follow-up. Kaplan- Meier analysis revealed that the OS was longer for patients with low RANKL expression than those with high RANKL expression (P=0.008, Figure 2A). No signiﬁcant association was found between RANK expression and OS (P=0.119, Figure 2B). We next divided patients into two groups according to the expression level of RANK and found that RANKL expression signiﬁcantly correlated with OS in the high RANK group (P=0.008, Figure 2C), while no signiﬁcant association was observed in the low RANK group (P=0.634, Figure 2D). In univariate analysis using COX proportional-hazard models, male, advanced pTNM stage, advanced T stage, presence of
As a receptor tyrosine kinase that plays essential roles in both normal physiological conditions and cancerous condi- tions, EGFR can affect many important characteristics of a cancer’s phenotype, including evasion of apoptosis, prolif- eration, invasion, and metastasis. In a previous study, the frequency of EGFR-positive tumors among young patients was found to be higher than among older women (5.9% vs. 3.3%, respectively), although the difference was not statis- tically significant. 16 Based on the results of that study and the data presented in this report, EGFR expression appears to play an important role in the early onset of breastcancer, and we hypothesize that this role is related to intrinsic genetic differences that result in an adverse outcome that is associated with the aggressive breast carcinomas. Further- more, we propose that EGFR immunostaining and/or amplification should be further investigated as predictor of a patient’s response to targeted therapy.
Evaluation of the association between REG4 expression and clinicopathological parameters or MUC expressions was done by the exact Pearson chi-square test or the exact linear-by-linear association test for ordered parameters. Disease-specific overall survival was counted from date of surgery to date of death from colorectal cancer, or until end of follow-up. Survival analysis by the Kaplan-Meier method was compared by the log rank test. The Cox regression proportional hazard model served for uni- and multivariable survival analysis, adjusted for sex, age, Dukes classification, and differentiation. Testing of the Cox model assumption of constant hazard ratios over time involved the inclusion of a time-dependent covariate separately for each testable variable. The hazard ratio of differentiation was analyzed in two periods (0 to 1.25 and 1.25 to 5 years) in order to meet the assumptions of the Cox model, with the time-dependent Cox model. Interaction terms were considered and an interaction between REG4 expression and age emerged. We therefore calculated the prognostic role of REG4 separately for patients under and over 65. All tests were two-sided. A p-value of 0.05 was considered significant. All statistical analyses were done with SPSS version 20.0 (IBM SPSS Statistics, version 20.0 for Mac; SPSS, Inc., Chicago, IL).
SK4 channels are expressed functionally inbreastcancer cell lines Four well-characterized breastcancer cell lines, MCF-7, T47D, MDA-MB-231 (MDA-231) and MDA-MB-468 (MDA-468), were included. We first found that SK4 proteins were expressed in MCF-7, MDA-468 and MDA-231 cells but showed little expression in T47D cells (Fig 2A). At the same time, we detected the EMT state of the cell lines and found that MCF-7, T47D and MDA-468 cells expressed the epithelial cell marker E-cadherin but did not express the mesen- chymal cell marker Vimentin, whereas MDA-231 cells exhibited the opposite expression pat- tern (Fig 2A). Next, we measured the expression levels of SK4 mRNA inbreastcancer cells. SK4 mRNA was detected in all cell lines (Fig 2B). With MCF-7 cells as the control, the relative SK4 mRNA levels were 1.00 ± 0.06 times in MCF-7 cells, 0.06 ± 0.01 in T47D cells, 10.38 ± 1.09 in MDA-468 cells, and 3.97 ± 0.27 in MDA-231 cells. Again, we investigated the expression of ER in the T47D, MDA-231 and MDA-468 cell lines. Fig 2C shows a clear and specific band in T47D cells but not in MDA-468 or MDA-231 cells, indicating that ER protein was only expressed in T47D cells. We also located the SK4 proteins in these four cell lines. In Fig 2D–2F, SK4 proteins (red color) were clearly detected in these 3 cell lines with an enriched signal on the cell membrane, while the signal was relatively negativein T47D cells (Fig 2G). These data suggested that SK4 protein was expressed in MCF-7, MDA-468 and MDA-231 cells but showed little expression in T47D cells.
A selective inhibitor of 20-HETE synthesis, HET0016, has been reported to inhibit angiogenesis. 20-HETE has been known as a second mitogenic messenger of angiogenesis inducing growth factors. HET0016 effects were analyzed on MDA- MB-231 derived breastcancerin mouse and in vitro cell line. MDA-MB-231 tumor cells were implanted in animals’ right flank and randomly assigned to early (1 and 2), starting treatments on day 0, or delayed groups (3 and 4) on day 8 after implantation of tumor. Animals received HET0016 (10 mg/kg) treatment via intraperitoneal injection for 5 days/week for either 3 or 4 weeks. Control group received vehicle treatment. Tumor sizes were measured on days 7, 14, 21, and 28 and the animals were euthanized on day 22 and 29. Proteins were extracted from the whole tumor and from cells treated with 10 mM HET0016 for 4 and 24 hrs. Protein array kits of 20 different cytokines/factors were used. ELISA was performed to observe the HIF-1a and MMP-2 protein expression. Other markers were confirmed by IHC. HET0016 significantly inhibited tumor growth in all treatment groups at all-time points compared to control (p,0.05). Tumor growth was completely inhibited on three of ten animals on early treatment group. Treatment groups showed significantly lower expression of pro-angiogenic factors compared to control at 21 days; however, there was no significant difference in HIF-1a expression after treatments. Similar results were found in vitro at 24 hrs of HET0016 treatment. After 28 days, significant increase of angiogenin, angiopoietin-
Postoperative wound infection in an instrumented spine patient is often disastrous. Management includes implant removal leading to spine instability. Negative pressure wound therapy (NPWT) applied to the spine surgical wound is one of the wound care technique with successful results. We report a case of a man who sustained Chance fracture of Lumbar 1 (L1) vertebra treated with long segment posterior instrumentation, who unfortunately developed Extended-spectrum beta-lactamase (ESBL) positive E. coli infection one month after the operation. After careful debridement of the wound, the implant became exposed. Three cycles of NPWT were applied and the wound healed with granulation tissue completely covering the implant, and thus negating the need to remove the implant. In conclusion, the NPWT is a good alternative in postoperative wound management especially in an instrumented spine patient.
The chemotherapy regimen was then changed to a combination of dexamethasone, thalidomide, doxorubicin, cisplatin, cyclophosphamide, and etoposide (DT-PACE). After three cycles of this chemotherapeutic regimen, the mono- colonal component reduced completely and there was a reduction in the extramedullary plasmacytomas (breast, ster- num, and CNS). Treatment then proceeded with high-dose melphalan and rescue with autologous hematopoietic progen- itor stem cell transplantation and subsequent radiotherapy against the sternal plasmocytoma.
was aromatase inhibitors). For adjuvant therapy, a total of 180 (5β.γ%) of γ44 patients were treated with hormonal therapy, γ6 (10.4%) with chemotherapy which was FEC with or without (w/o) DOC or AC w/o PTX, 81 (βγ.5%) patients were treated with both hormonal and chemo-therapy, respectively. In addition, 18 patients were treated with targeted therapy using trastuzumab simultaneously. No therapy was administered to γ5 patients, and no detailed information on therapy could be obtained for 1β patients. When tumor recurs, patients with hormone receptor-positive tumors and non-visceral metastases were treated with endocrine therapy, such as antiestrogens, aromatase inhibitors, and medroxyprogesterone acetate. For patients with Herβ overexpressed tumors, tastuzumab was applied. Other patients were treated with chemotherapy such as anthracycline containing regimens, taxanes, capecitabine, and vinorelbine. Patients were periodically examined at the Kumamoto University Hospital or affiliated hospitals. The patients were observed every γ months for 5 years and every 1 year thereafter. Recurrence was defined when positive spots were found by physical examination and/or by imaging diagnosis during follow-up period. The median follow-up period was 66.5 months (range, 0.βγ - 145 months).